Immobilization of alkaline protease produced by Streptomyces rochei strain NAM-19 in solid state fermentation based on medium optimization using central composite design

This study evaluated Streptomyces rochei strain NAM-19 solid-state fermentation of agricultural wastes to produce alkaline protease. Alkaline protease production increased with flaxseed, rice bran, and cheese whey fermentation reaching 147 U/mL at 48 h. Statistical optimization of alkaline protease...

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Bibliographic Details
Published in:3 Biotech 2024-06, Vol.14 (6), p.161, Article 161
Main Authors: El-Shazly, Asmaa I., Wahba, Marwa I., Abdelwahed, Nayera A. M., Shehata, Abeer N.
Format: Article
Language:English
Subjects:
Agricultural wastes
Agriculture
Alkaline protease
Ammonium sulfate
Azocasein
Bioinformatics
Biomaterials
Biopolymers
Biotechnology
Cancer Research
Carrageenan
Casein
Cheese
Chemistry
Chemistry and Materials Science
Design optimization
Enzymes
Fermentation
Fractionation
Gelatin
Immobilization
Immobilized enzymes
Inoculum
Metal ions
Optimization
Original Article
Protease
Solid state
Solid state fermentation
Stem Cells
Streptomyces rochei
Thermal stability
Whey
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Summary:This study evaluated Streptomyces rochei strain NAM-19 solid-state fermentation of agricultural wastes to produce alkaline protease. Alkaline protease production increased with flaxseed, rice bran, and cheese whey fermentation reaching 147 U/mL at 48 h. Statistical optimization of alkaline protease production was performed using the central composite design (CDD). Results of CDD and the optimization plot showed that 4.59 g/L flaxseed, 4.31 g/L rice bran, 4.17 mL cheese whey, and a vegetative inoculum size of 7.0% increased alkaline protease production by 27.2% reaching 186 U/mL. Using the 20–70% ammonium sulfate fractionation method, the optimally produced enzyme was partially purified to fivefold. The partially purified alkaline protease was then covalently immobilized on a biopolymer carrier, glutaraldehyde-polyethylene-imine-κ-carrageenan (GA-PEI-Carr), with 90% immobilization efficiency. Characterizations revealed that immobilization improved thermostability, reusability, optimum temperature, and sensitivity towards metal ions of the free enzyme. The optimal temperature for free and immobilized enzymes was 40 and 50 °C, respectively. Both enzymes had the same optimum pH of 10. Immobilization increased K m from 19.73 to 26.52 mM and V max from 56.7 to 62.5 mmol min −1 L −1 . The immobilized enzyme retained 35% of its initial activity at 70 °C, while the free enzyme retained only 5%. The immobilized enzyme kept 80% of its initial activity at the 20th cycle. After 7 weeks of storage, the free enzyme lost all its initial activity, whereas the immobilized enzyme retained 50%. The free and immobilized enzymes were able to hydrolyze gelatin, and azo-casein demonstrating different relative activity, 85, 80, 90 and 95%, respectively, compared to casein (100%).
ISSN:2190-572X
2190-5738
DOI:10.1007/s13205-024-04003-9